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. 2020:2124:251-261.
doi: 10.1007/978-1-0716-0356-7_13.

Biolistic DNA Delivery in Turfgrass Embryonic Callus Initiated from Mature Seeds

Man Zhou  1 , Junming Zhao  2 , Dayong Li  2 , Shuangrong Yuan  2 , Ning Yuan  2 , Zhigang Li  2 , Haiyan Jia  2 , Fangyuan Gao  2 , Bekir San  2 , Qian Hu  2 , Hong Luo  3
Affiliations

Biolistic DNA Delivery in Turfgrass Embryonic Callus Initiated from Mature Seeds

Man Zhou et al. Methods Mol Biol. 2020.

Abstract

We describe a protocol for the establishment and preparation of creeping bentgrass (Agrostis stolonifera L.) cultivar "Penn A-4" embryonic calli, biolistic transformation, selection, and regeneration of transgenic plants. The embryonic callus is initiated from mature seeds, maintained by visual selection under the dissecting microscope and subjected to bombardment with plasmid DNA containing a bialaphos-resistance (bar) gene. PCR, Southern, and Northern blot analyses are used to confirm the transgene integration and expression.

Keywords: Biolistic; DNA delivery; Embryonic callus; Gene gun; Genetic transformation; Mature seed; Turfgrass.

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